期刊
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY
卷 457, 期 2, 页码 561-568出版社
SPRINGER
DOI: 10.1007/s00424-008-0539-9
关键词
Dye labeling; Neurobiotin; Electroporation; Lucifer yellow; Dye coupling; Retina
类别
资金
- National Health and Medical Research Council (Australia)
- Australian Research Council Centre of Excellence in Vision Science
Intracellular sharp-electrode, whole-cell patch clamp and juxtacellular labeling methods have previously been developed for combined analysis of neuronal structure and function. We describe a novel electroporation technique for labeling neurons with Neurobiotin, using patch electrodes in a semi-loose seal configuration (R=100-300 MO) with very small amplitude pulses (50 mV). The addition of 2% Neurobiotin to the intracellular solution in the patch electrode reduces the dielectric membrane breakdown voltage threshold by about threefold. The resulting pore formation allows for (1) the stable recording of spontaneous and light-evoked postsynaptic potentials without significant cytoplasmic washout and (2) the passage of dye without spillover. The efficiency and reliability of the method makes it particularly suitable for the serial recording and labeling of multiple neurons in a small area of tissue.
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